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Related Experiment Videos

A new covalently modified support for gas-liquid phase sequencing.

R Frank, K Ashman

    Biological Chemistry Hoppe-Seyler
    |July 1, 1986
    PubMed
    Summary
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    A novel method activates glass fiber supports for protein and peptide immobilization, improving detection sensitivity in gas-liquid phase sequencing. This new method enhances yields and reduces background noise compared to polybrene.

    Area of Science:

    • Biochemistry
    • Analytical Chemistry
    • Materials Science

    Background:

    • Current methods for immobilizing proteins and peptides for gas-liquid phase sequencing often require precycling and can suffer from high background contamination.
    • Polybrene is a commonly used carrier, but it has limitations that can be overcome with alternative support materials.

    Purpose of the Study:

    • To describe a new method for activating glass fiber supports for protein and peptide immobilization.
    • To evaluate the advantages of this new support over existing carriers like polybrene.
    • To enhance detection sensitivity in gas-liquid phase sequencing.

    Main Methods:

    • Glass fibers were activated using an acid treatment.
    • Covalent coating of glass fibers with quaternary ammonium groups was performed.

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  • The activated glass supports were used for immobilizing proteins and peptides.

    • Main Results:

    • The new glass fiber support requires no precycling, unlike polybrene.
    • Initial yields of immobilized proteins and peptides were improved.
    • Background contamination was significantly lower, leading to increased detection sensitivity.
    • The activated glass demonstrated high binding capacity for both proteins and peptides.

    Conclusions:

    • The developed acid activation and covalent coating method provides an effective way to prepare glass fiber supports for protein and peptide immobilization.
    • This new method offers significant advantages over polybrene, including no precycling, improved yields, and lower background.
    • The activated glass supports are suitable for sequencing proteins and peptides isolated by HPLC and electroelution from polyacrylamide gels, enhancing overall detection sensitivity.